Cellular hybridization for BDNF. trkB, and NGF mRNAs and BDNF-immunoreactivity in rat forebrain after pilocarpine-induced status epilepticus
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Abstract
The messenger RNAs (mRNAs) for the neurotrophins, brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF), are upregulated during epileptic seizure activity, as visualized by in situ hybridization techniques. Neurotrophins might be protective against excitotoxic cell stress, and the upregulation during seizures might provide such cell protection. In this study, a high dose of pilocarpine (300 mg/kg) was used to induce long-lasting, limbic motor status epilepticus and a selective pattern of brain damage. The regulation of BDNF, trkB, and NGF mRNA was studied by in situ hybridization at 1, 3, 6, and 24 h after induction of limbic motor status epilepticus. BDNF immunoreactivity was examined with an anti-peptide antibody and the neuropathological process studied in parallel. BDNF mRNA increased in hippocampus, neocortex, piriform cortex, striatum, and thalamus with a maximum at 3–6 h. Hybridization levels increased earlier in the resistant granule and CA1 cells as compared to the vulnerable CA3 neurons. BDNF immunoreactivity was elevated in dentate gyrus at 3–6 h. trKB mRNA increased in the entire hippocampus. NGF mRNA in hippocampus appeared in dentate gyrus at 3–6 h and declined in hilar neurons at 6–24 h. Cell damage was found in the CA3 area, entire basal cortex, and layers II/III of neocortex. Endogenous neurotrophins are upregulated during status epilepticus caused by pilocarpine, which is related to the coupling between neuronal excitation and trophic factor expression. This upregulation of neurotrophic factors may serve endogenous protective effects; however, the excessive levels of neuronal hyperexcitation resulting from pilocarpine seizures lead to cell damage which cannot be prevented by endogenous neurotrophins.
Key words
Neurotrophins BDNF In situ hybridization Immunohistochemistry Status epilepticus RatPreview
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References
- Ballarín M, Ernfors P, Lindefors N, Persson H (1991) Hippocampal damage and kainic acid injection induce a rapid increase in mRNA for BDNF and NGF in the rat brain. Exp Neurol 114: 35–43Google Scholar
- Barde Y-A (1989) Trophic factors and neuronal survival. Neuron 2: 1525–1534CrossRefPubMedGoogle Scholar
- Beck KD, Lamballe F, Klein R, Barbacid M, Schauwecker PE, McNeill TH, Finch CE, Hefti F, Day JR (1993) Induction of noncatalytic trkB neurotrophin receptors during axonal sprouting in the adult hippocampus. J Neurosci 13: 4001–4014Google Scholar
- Beck T, Wree A, Sauer DD (1992) Chronic infusion of nerve growth factor does not rescue pyramidal cells after transient forebrain ischemia in the rat. Neurosci Lett 135: 252–254Google Scholar
- Bengzon J, Söderström S, Kokaia Z, Kokaia M, Ernfors P, Persson H, Ebendahl T, Lindvall O (1992) Widespread increase of nerve growth factor protein in the rat forebrain after kindling-induced seizures. Brain Res 587: 338–342Google Scholar
- Berkemeier LR, Winslow JW, Kaplan DR, Nikolics K, Goeddel DV, Rosenthal A (1991) Neurotrophin-5: A novel neurotrophic factor that activates trk and trkB. Neuron 7: 857–866CrossRefPubMedGoogle Scholar
- Berninger B, Garcia DE, Inagaki N, Hahnel C, Lindholm D (1993) BDNF and NT-3 induce intracellular Ca2+ elevation in hippocampal neurones. NeuroReport 4: 1303–1306Google Scholar
- Berzaghi MP, Cooper J, Castrén E, Zafra F, Sofroniew M, Thoenen H, Lindholm D (1993) Cholinergic regulation of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) but not neurotrophin-3 (NT-3) mRNA levels in developing hippocampus. J Neurosci 13: 3818–3826Google Scholar
- Boatell LL, Lindefors N, Ballarín M, Ernfors P, Mahy N, Persson H (1992) Activation of basal forebrain cholinergic neurons differentially regulates brain-derived neurotrophic factor mRNA expression in different projection areas. Neurosci Lett 136: 203–208Google Scholar
- Castrén E, Zafra F, Thoenen H, Lindholm D (1992) Light regulates expression of brain-derived neurotrophic factor mRNA in rat visual cortex. Proc Natl Acad Sci 89: 9444–9448Google Scholar
- Castrén E, Berzaghi MP, Lindholm D, Thoenen H (1993) Differential effects of MK-801 on brain-derived neurotrophic factor mRNA levels in different regions of the rat brain. Exp Neurol 122: 244–252Google Scholar
- Ceccatelli S, Ernfors P, Villar MJ, Persson H, Hökfelt T (1991) Expanded distribution of mRNA for nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3 in the rat brain after colchicine treatment. Proc Natl Acad Sci 88: 10352–10356Google Scholar
- Cheng B, Mattson MP (1991) NGF and bFGF protect rat hippocampal and human cortical neurons against hypoglycemie damage by stabilizing calcium homeostasis. Neuron 7: 1031–1041Google Scholar
- Choi DW (1988) Glutamate neurotoxicity and diseases of the central nervous system. Neuron 1: 623–634PubMedGoogle Scholar
- Clifford DB, Olney JW, Maniotis A, Collins RC, Zorumski CF (1987) The functional anatomy and pathology of lithium-pilocarpine and high dose pilocarpine seizures. Neuroscience 23: 953–968Google Scholar
- Comelli MC, Seren MS, Guidolin D, Manev RM, Favaron M, Rimland JM, Canella R, Negro A, Manev H (1992) Photochemical stroke and brain-derived neurotrophic factor (BDNF) mRNA expression. NeuroReport 3: 473–476Google Scholar
- Davies SW, Beardsall K (1992) Nerve growth factor selectively prevents excitotoxin induced degeneration of striatal cholinergic neurons. Neurosci Lett 140: 161–164Google Scholar
- Dugich-Djordjevic MM, Tocco G, Lapchak PA, Pasinetti GM, Najm I, Baudry M, Hefti F (1992) Regionally specific and rapid increases in brain-derived neurotrophic factor messenger RNA in the adult rat brain following seizures induced by systemic administration of kainic acid. Neuroscience 47: 303–315Google Scholar
- Ernfors P, Wetmore C, Olson L, Persson H (1990) Identification of cells in rat brain and peripheral tissues expressing mRNA for members of the nerve growth factor family. Neuron 5: 511–526Google Scholar
- Ernfors P, Bengzon J, Kokaia Z, Persson H, Lindvall O (1991) Increased levels of messenger RNAs for neurotrophic factors in the brain during kindling epileptogenesis. Neuron 7: 165–176Google Scholar
- Frim DM, Yee WM, Isacson O (1993) NGF reduces striatal excitotoxic neuronal loss without affecting concurrent neuronal stress. NeuroReport 4: 655–658Google Scholar
- Gall CM (1993) Seizure-induced changes in neurotrophin expression: implications for epilepsy. Exp Neurol 124: 150–166Google Scholar
- Gall CM, Isackson PJ (1989) Limbic seizures increase neuronal production of messenger RNA for nerve growth factor. Science 245: 758–761Google Scholar
- Gall C, Murray K, Isackson PJ (1991) Kainic acid-induced seizures stimulate expression of nerve growth factor mRNA in rat hippocampus. Mol Brain Res 9: 113–123Google Scholar
- Gwag BJ, Springer JE (1993) Activation of NMDA receptors increases brain-derived neurotrophic factor (BDNF) mRNA expression in the hippocampal formation. NeuroReport 5: 125–128Google Scholar
- Hantzopoulos PA, Suri C, Glass DJ, Goldfarb MP, Yancopoulos GD (1994) The low affinity NGF receptor, p75, can collaborate with each of the trks to potentiate functional responses to the neurotrophins. Neuron 13: 187–201Google Scholar
- Hofer M, Pagliusi SR, Hohn A, Leibrock J, Barde YA (1990) Regional distribution of brain-derived neurotrophic factor mRNA in the adult mouse brain. EMBO J 9: 2459–2464PubMedGoogle Scholar
- Humpel C, Wetmore C, Olson L (1993) Regulation of brain-derived neurotrophic factor messenger RNA and protein at the cellular level in pentylenetetrazol-induced epileptic seizures. Neuroscience 53: 909–918Google Scholar
- Ingvar M, Folbergrova J, Siesjö BK (1987) Metabolic alterations underlying the development of hypermetabolic necrosis in the substantia nigra in status epilepticus. J Cereb Blood Flow Metab 7: 103–108Google Scholar
- Isackson PJ, Huntsman MM, Murray KD, Gall CM (1991) BDNF mRNA expression is increased in adult rat forebrain after limbic seizures: temporal patterns of induction distinct from NGF. Neuron 6: 937–948Google Scholar
- Kiessling M, Xie Y, Kleihues P (1984) Regional impairment of protein synthesis in the rat brain during bicuculline-induced seizures. Brain Res 296: 1–13Google Scholar
- Klein R, Martin-Zanca D, Barbacid M, Parada LF (1990) Expression of the tyrosine kinase receptor gene trkB is confined to the murine embryonic and adult nervous system. Development 109: 845–850Google Scholar
- Kokaia Z, Gidö G, Ringstedt T, Bengzon J, Kokaia M, Siesjö BK, Persson H, Lindvall O (1993) Rapid increase of BDNF mRNA levels in cortical neurons following spreading depression: regulation by glutamatergic mechanisms independent of seizure activity. Mol Brain Res 19: 277–286Google Scholar
- Lauterborn JC, Tran TMD, Isackson PJ, Gall CM (1993) Nerve growth factor mRNA is expressed by GABAergic neurons in rat hippocampus. NeuroReport 5: 273–276Google Scholar
- Lauterborn JC, Isackson PJ, Gall CM (1994) Seizure-induced increases in NGF mRNA exhibit different time courses across forebrain regions and are biphasic in hippocampus. Exp Neurol 125: 22–40Google Scholar
- Leibrock J, Lottspeich F, Hohn A, Hofer M, Hengerer B, Masiakowski P, Thoenen H, Barde YA (1989) Molecular cloning and expression of brain-derived neurotrophic factor. Nature 341: 149–152CrossRefPubMedGoogle Scholar
- Lindefors N, Ernfors P, Falkenberg T, Persson H (1992) Septal cholinergic afferents regulate expression of brain-derived neurotrophic factor and β-nerve growth factor mRNA in rat hippocampus. Exp Brain Res 88: 78–90Google Scholar
- Lindsay RM (1993) Brain-derived neurotrophic factor: an NGFrelated neurotrophin. In: Loughlin SE, Fallon JH (eds) Neurotrophic factors. Academic, San Diego, pp 257–284Google Scholar
- Lindsay RM, Wiegand SJ, Altar CA, DiStefano PS (1994) Neurotrophic factors: from molecule to man. Trends Neurosci 17: 182–190Google Scholar
- Lindvall O, Ernfors P, Bengzon J, Kokaia Z, Smith M-L, Siesjö BK, Persson H (1992) Differential regulation of mRNAs for nerve growth factor, brain derived neurotrophic factor, and neurotrophin 3 in the adult rat brain following cerebral ischemia and hypoglycemie coma. Proc Natl Acad Sci 89: 648–652Google Scholar
- Liu Z, D'Amore PA, Mikati M, Gatt A, Holmes GL (1993) Neuroprotective effect of chronic infusion of basic fibroblast growth factor on seizure-associated hippocampal damage. Brain Res 626: 335–338Google Scholar
- Longo FM, Holtzman DM, Grimes ML, Mobley WC (1993) Nerve growth factor: actions in the peripheral and central nervous systems. In: Loughlin SE, Fallon JH (eds) Neurotrophic factors. Academic, San Diego, pp 209–256Google Scholar
- Loughlin SE, Fallon JH (1993) Neurotrophic factors. Academic, SanDiegoGoogle Scholar
- Meldrum BS (1991) Excitotoxicity and epileptic brain damage. Epilepsy Res 10: 55–61Google Scholar
- Merlio J-P, Ernfors P, Jaber M, Persson H (1992) Molecular cloning of rat trkC and distribution of cells expressing messenger RNAs for members of the trk family in the rat central nervous system. Neuroscience 51: 513–532CrossRefPubMedGoogle Scholar
- Merlio J-P, Ernfors P, Kokaia Z, Middlemas DS, Bengzon J, Kokaia M, Smith M-L, Siesjö BK, Hunter T, Lindvall O, Persson H (1993) Increased production of the trkB protein tyrosine kinase receptor after brain insults. Neuron 10: 151–164Google Scholar
- Metsis M, Timmusk T, Arenas E, Persson H (1993) Differential usage of multiple brain-derived neurotrophic factor promoters in the rat brain following neuronal activation. Proc Natl Acad Sci 90: 8802–8806Google Scholar
- Middlemas DS, Lindberg RA, Hunter T (1991) TrkB, a neural receptor protein-tyrosine kinase: evidence for a full-length and two truncated receptors. Mol Cell Biol 11: 143–153Google Scholar
- Millan MH, Chapman AG, Meldrum BS (1993) Extracellular amino acid levels in hippocampus during pilocarpine-induced seizures. Epilepsy Res 14: 139–148Google Scholar
- Nawa H, Carnahan J, Gall C (1995) BDNF protein measured by a novel enzyme immunoassay in normal brain and after seizure: partial disagreement with mRNA levels. Eur J Neurosci 7: 1527–1535Google Scholar
- Phillips HS, Hains JM, Laramee GR, Rosenthal A, Winslow JW (1990) Widespread expression of BDNF but not NT 3 by target areas of basal forebrain cholinergic neurons. Science 250: 290–294Google Scholar
- Radeke MJ, Misko TP, Hsu C, Herzenberg LA, Shooter EM (1987) Gene transfer and molecular cloning of the rat nerve growth factor receptor. Nature 325: 593–597Google Scholar
- Rocamora N, Palacios JM, Mengod G (1992) Limbic seizures induce a differential regulation of the expression of nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3, in the rat hippocampus. Mol Brain Res 13: 27–33Google Scholar
- Rodriguez-Tebar A, Dechant G, Barde Y (1990) Binding of brain-derived neurotrophic factor to the nerve growth factor receptor. Neuron 4: 487–492Google Scholar
- Schmidt-Kastner R, Ingvar M (1994) Loss of immunoreactivity for glial fibrillary acidic protein (GFAP) in astrocytes as a marker for profound tissue damage in substantia nigra and basal cortical areas after status epilepticus induced by pilocarpine in rat. Glia 12: 165–172Google Scholar
- Schmidt-Kastner R, Ingvar M (1996) Laminar damage of neurons and astrocytes in neocortex and hippocampus of rat after long-lasting status epilepticus induced by pilocarpine. Epilepsy Res [Suppl] (in press)Google Scholar
- Schmidt-Kastner R, Heim C, Sontag KI-H (1991) Damage of substantia nigra pars reticulata during pilocarpine-induced status epilepticus in the rat: immuno-histochemical study of neurons, astrocytes and serum-protein extravasation. Exp Brain Res 86: 125–140Google Scholar
- Schmidt-Kastner R, Humpel C, Wetmore C, Ingvar M, Olson L (1993) Neurotrophins and neuropathology after pilocarpine-induced status epilepticus. Soc Neurosci Abstr 19: 606Google Scholar
- Shigeno A, Mima T, Takakura K, Graham DI, Kato G, Hashimoto Y, Furukawa S (1991) Amelioration of delayed neuronal death in the hippocampus by nerve growth factor. J Neurosci 11: 2914–2919Google Scholar
- Shozuhara H, Onodera H, Katoh-Semba R, Kato K, Yamasaki Y, Kogure K (1992) Temporal profiles of nerve growth factor β-subunit level in rat brain regions after transient ischemia. J Neurochem 59: 175–180Google Scholar
- Soppet D, Escandon E, Maragos J, Kaplan DR, Hunter T, Nikolics K, Parada LF (1991) The neurotrophic factors BDNF and NT-3 are ligands for the trkB tyrosine kinase receptor. Cell 65: 895–903Google Scholar
- Squinto SP, Stitt TN, Aldrich TH, Davis SM, Bianco SM, Radziejewski C, Glass DJ, Masiakowski P, Furth ME, Valenzuela DM, DiStefano PS, Yancopoulos CD (1991) trkB encodes a functional receptor for brain-derived neurotrophic factor and neurotrophin-3 but not for NGF. Cell 65: 885–893Google Scholar
- Thoenen H (1991) The changing scene of neurotrophic factors. Trends Neurosci 14: 165–170CrossRefPubMedGoogle Scholar
- Timmusk T (1994) Structure and regulation of neurotrophins and their low affinity receptor genes. Ph.D.Thesis, Karolinska Institute, StockholmGoogle Scholar
- Turski L, Meldrum, BS, Cavalheiro EA, Calderazzo-Filho LS, Bortolotto A, Ikonomidou-Turski C, Turski WA (1989) Paradoxical anticonvulsant activity of the excitatory amino acid N-methyl-D-aspartate in the rat caudate-putamen. Proc Natl Acad Sci 86: 1694–1697Google Scholar
- Turski WA, Cavalheiro EA, Schwarz M, Czuczwar SJ, Kleinrok Z, Turski L (1983) Limbic seizures produced by pilocarpine in rats: behavioural, electroencephalographic and neuropathological study. Behav Brain Res 9: 315–335Google Scholar
- Wetmore C, Ernfors P, Persson H, Olson L (1990) Localization of brain-derived neurotrophic factor mRNA to neurons in the brain by in situ hybridization. Exp Neurol 109: 141–152Google Scholar
- Wetmore C, Cao Y, Petterson RF, Olson L (1991) Brain-derived trophic factor: subcellular compartmentalization and interneuronal transfer as visualized by anti-peptide antibodies. Proc Natl Acad Sci 88: 9843–9846Google Scholar
- Wetmore C, Cao Y, Petterson RF, Oison L (1993) Brain-derived neurotrophic factor (BDNF) peptide antibodies: characterization using a vaccinia virus expression system. J Histochem Cytochem 41: 521–533Google Scholar
- Wetmore C, Olson L, Bean AJ (1994) Regulation of brain-derived neurotrophic factor (BDNF) expression and release from hippocampal neurons is mediated by non-NMDA type glutamate receptors. J Neurosci 14: 1688–1700Google Scholar
- Whittemore SR, Friedman PL, Larhammar D, Persson H, Gonzalez CM, Holets VR (1988) Rat β-nerve growth factor sequence and site of synthesis in the adult hippocampus. J Neurosci Res 20: 403–410Google Scholar
- Yamamoto S, Yoshimine T, Fujita T, Kuroda R, Irie T, Fujioka K, Hayakawa T (1992) Protective effect of NGF atecollagen mini-pellet on the hippocampal delayed neuronal death in gerbils. Neurosci Lett 141: 161–165Google Scholar
- Yan Q, Matheson CR, Lopez OT, Hawkins N, Rosenfeld RD, Bennett L (1994) Immunohistochemical study of the expression of brain-derived neurotrophic factor (BDNF) in adult rat brain. Soc Neurosci Abstr 20: 1306Google Scholar
- Zafra F, Hengerer B, Leibrock J, Thoenen H, Lindholm D (1990) Activity dependent regulation of BDNF and NGF mRNAs in the rat hippocampus is mediated by non-NMDA glutamate receptors. EMBO J 9: 3545–3550PubMedGoogle Scholar
- Zafra F, Castrén E, Thoenen H, Lindholm D (1991) Interplay between glutamate and γ-aminobutyric acid transmitter systems in the physiological regulation of brain-derived neurotrophic factor and nerve growth factor synthesis in hippocampal neurons. Proc Natl Acad Sci 88: 10037–10041Google Scholar
- Zafra F, Lindholm D, Castrén E, Hartikka J, Thoenen H (1992) Regulation of brain-derived neurotrophic factor and nerve growth factor mRNA in primary cultures of hippocampal neurons and astrocytes. J Neurosci 12: 4793–4799Google Scholar